<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2025-12-12</periodOfReport>

    <issuer>
        <issuerCik>0001641281</issuerCik>
        <issuerName>Bolt Biotherapeutics, Inc.</issuerName>
        <issuerTradingSymbol>BOLT</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0002023538</rptOwnerCik>
            <rptOwnerName>Nemec Sarah</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>900 CHESAPEAKE DRIVE</rptOwnerStreet1>
            <rptOwnerStreet2></rptOwnerStreet2>
            <rptOwnerCity>REDWOOD CITY</rptOwnerCity>
            <rptOwnerState>CA</rptOwnerState>
            <rptOwnerZipCode>94063</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>false</isDirector>
            <isOfficer>true</isOfficer>
            <isTenPercentOwner>false</isTenPercentOwner>
            <isOther>false</isOther>
            <officerTitle>Senior VP, Finance and PAO</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>false</aff10b5One>

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                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>4499</value>
                </transactionShares>
                <transactionPricePerShare>
                    <footnoteId id="F11"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F1"/>
            </exerciseDate>
            <expirationDate>
                <value>2034-07-22</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>4499</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>0</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
        <derivativeTransaction>
            <securityTitle>
                <value>Employee Stock Option (Right to Buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>5.44</value>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2025-12-12</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>11299</value>
                </transactionShares>
                <transactionPricePerShare>
                    <footnoteId id="F13"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F12"/>
            </exerciseDate>
            <expirationDate>
                <value>2035-04-30</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>11299</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>11299</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
        <derivativeTransaction>
            <securityTitle>
                <value>Employee Stock Option (Right to Buy)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>7.02</value>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2025-12-12</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>D</transactionCode>
                <equitySwapInvolved>false</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>11299</value>
                </transactionShares>
                <transactionPricePerShare>
                    <footnoteId id="F13"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>D</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F12"/>
            </exerciseDate>
            <expirationDate>
                <value>2035-04-30</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>11299</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>0</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">The option is fully vested and exercisable.</footnote>
        <footnote id="F2">The reporting person agreed to cancellation of an option granted on September 3, 2020, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F3">The reporting person agreed to cancellation of an option granted on February 4, 2021, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F4">The reporting person agreed to cancellation of an option granted on December 22, 2021, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F5">The reporting person agreed to cancellation of an option granted on February 18, 2022, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F6">For so long as the Reporting Person continuously provides services to the Issuer this option will vest with respect to the shares as follows: 1/36th of the Shares will vest upon the Reporting Person completing each month of continuous service following January 1, 2023.</footnote>
        <footnote id="F7">The reporting person agreed to cancellation of an option granted on January 19, 2023, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F8">For so long as the Reporting Person continuously provides services to the Issuer this option will vest with respect to the shares as follows: 1/36th of the Shares will vest upon the Reporting Person completing each month of continuous service following January 1, 2024.</footnote>
        <footnote id="F9">The reporting person agreed to cancellation of an option granted on March 4, 2024, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F10">For so long as the Reporting Person continuously provides services to the Issuer this option will vest with respect to the shares as follows: 1/36th of the Shares will vest upon the Reporting Person completing each month of continuous service following July 15, 2024.</footnote>
        <footnote id="F11">The reporting person agreed to cancellation of an option granted on July 23, 2024, in exchange for a new option having a lower exercise price.</footnote>
        <footnote id="F12">For so long as the Reporting Person continuously provides services to the Issuer this option will vest with respect to the shares as follows: 1/36th of the Shares will vest upon the Reporting Person completing each month of continuous service following January 1, 2025.</footnote>
        <footnote id="F13">The reporting person agreed to cancellation of an option granted on May 1, 2025, in exchange for a new option having a lower exercise price.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ William P. Quinn, Attorney-in-Fact</signatureName>
        <signatureDate>2025-12-16</signatureDate>
    </ownerSignature>
</ownershipDocument>
